Tocilizumab in refractory Caucasian Takayasu's arteritis: a multicenter study of 54 patients and literature review

Objective: To assess the efficacy and safety of tocilizumab (TCZ) in Caucasian patients with refractory Takayasu's arteritis (TAK) in clinical practice. Methods: A multicenter study of Caucasian patients with refractory TAK who received TCZ. The outcome variables were remission, glucocorticoid-sparing effect, improvement in imaging techniques, and adverse events. A comparative study between patients who received TCZ as monotherapy (TCZMONO) and combined with conventional disease modifying anti-rheumatic drugs (cDMARDs) (TCZCOMBO) was performed. Results: The study comprised 54 patients (46 women/8 men) with a median [interquartile range (IQR)] age of 42.0 (32.5-50.5) years. TCZ was started after a median (IQR) of 12.0 (3.0-31.5) months since TAK diagnosis. Remission was achieved in 12/54 (22.2%), 19/49 (38.8%), 23/44 (52.3%), and 27/36 (75%) patients at 1, 3, 6, and 12 months, respectively. The prednisone dose was reduced from 30.0 mg/day (12.5-50.0) to 5.0 (0.0-5.6) mg/day at 12 months. An improvement in imaging findings was reported in 28 (73.7%) patients after a median (IQR) of 9.0 (6.0-14.0) months. Twenty-three (42.6%) patients were on TCZMONO and 31 (57.4%) on TCZCOMBO: MTX (n = 28), cyclosporine A (n = 2), azathioprine (n = 1). Patients on TCZCOMBO were younger [38.0 (27.0-46.0) versus 45.0 (38.0-57.0)] years; difference (diff) [95% confidence interval (CI) = -7.0 (-17.9, -0.56] with a trend to longer TAK duration [21.0 (6.0-38.0) versus 6.0 (1.0-23.0)] months; diff 95% CI = 15 (-8.9, 35.5), and higher c-reactive protein [2.4 (0.7-5.6) versus 1.3 (0.3-3.3)] mg/dl; diff 95% CI = 1.1 (-0.26, 2.99). Despite these differences, similar outcomes were observed in both groups (log rank p = 0.862). Relevant adverse events were reported in six (11.1%) patients, but only three developed severe events that required TCZ withdrawal. Conclusion: TCZ in monotherapy, or combined with cDMARDs, is effective and safe in patients with refractory TAK of Caucasian origin.Funding: This work was partially supported by RETICS Programs, RD08/0075 (RIER), RD12/0009/0013 and RD16/0012 from “Instituto de Salud Carlos III” (ISCIII) (Spain)

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7473G>A (p.=) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of 3 mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that 4 of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease.info:eu-repo/semantics/publishedVersio

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to the identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7437G>A) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of three mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that four of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease. clinicaltrials.gov identifier:02869074

Unraveling the effect of silent, intronic and missense mutations on VWF splicing : contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to the identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7437G>A) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of three mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that four of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease. identifier:02869074

Tree growth response to drought partially explains regional-scale growth and mortality patterns in Iberian forests

Tree-ring data has been widely used to inform about tree growth responses to drought at the individual scale, but less is known about how tree growth sensitivity to drought scales up driving changes in forest dynamics. Here, we related tree-ring growth chronologies and stand-level forest changes in basal area from two independent data sets to test if tree-ring responses to drought match stand forest dynamics (stand basal area growth, ingrowth, and mortality). We assessed if tree growth and changes in forest basal area covary as a function of spatial scale and tree taxa (gymnosperm or angiosperm). To this end, we compared a tree-ring network with stand data from the Spanish National Forest Inventory. We focused on the cumulative impact of drought on tree growth and demography in the period 1981–2005. Drought years were identified by the Standardized Precipitation Evapotranspiration Index, and their impacts on tree growth by quantifying tree-ring width reductions. We hypothesized that forests with greater drought impacts on tree growth will also show reduced stand basal area growth and ingrowth and enhanced mortality. This is expected to occur in forests dominated by gymnosperms on drought-prone regions. Cumulative growth reductions during dry years were higher in forests dominated by gymnosperms and presented a greater magnitude and spatial autocorrelation than for angiosperms. Cumulative drought-induced tree growth reductions and changes in forest basal area were related, but initial stand density and basal area were the main factors driving changes in basal area. In drought-prone gymnosperm forests, we observed that sites with greater growth reductions had lower stand basal area growth and greater mortality. Consequently, stand basal area, forest growth, and ingrowth in regions with large drought impacts was significantly lower than in regions less impacted by drought. Tree growth sensitivity to drought can be used as a predictor of gymnosperm demographic rates in terms of stand basal area growth and ingrowth at regional scales, but further studies may try to disentangle how initial stand density modulates such relationships. Drought-induced growth reductions and their cumulative impacts have strong potential to be used as early-warning indicators of regional forest vulnerability.This study was financially supported by Xunta de Galicia, Grant/Award Number PGIDIT06PXIB502262PR, GRC GI-1809; INIA, Grant/Award Number RTA2006-00117; CANOPEE, 2014-2020-FEDER funds, Spanish Science Ministry RTI2018-096884-B-C31, RTI2018-096884-B-C33, AGL2017-83828-C2-2R, RTI2018-096884-B-C3,1 and RTI2018-096884-B-C32 projects. Gabriel Sangüesa-Barreda was supported by a “Juan de la Cierva-Formación” grant from MINECO (FJCI 2016-30121). Antonio Gazol and Paloma Ruiz-Benito were supported by a project “2018 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation.” Ana-Maria Hereş was supported by the project PN-III-P1-1.1-TE-2019-1099 financed by the Romanian Ministry of Education and Research through UEFISCDI. Raúl Sánchez-Salguero was supported by VULBOS project (UPO-1263216, FEDER Funds, Andalusia Regional Government, Consejería de Economía, Conocimiento, Empresas y Universidad 2014-2020). Paloma Ruiz-Benito was supported by the Community of Madrid Region under the framework of the multi-year Agreement with the University of Alcalá (Stimulus to Excellence for Permanent University Professors, EPU-INV/2020/010) and the University of Alcalá “Ayudas para la realización de Proyectos para potenciar la Creación y Consolidación de Grupos de Investigación.” Andrea Hevia was supported by PinCaR project (UHU-1266324, FEDER Funds, Andalusia Regional Government, Consejería de Economía, Conocimiento, Empresas y Universidad 2014-2020).Peer reviewe

Molecular and clinical profile of von Willebrand disease in Spain (PCM-EVW-ES) : comprehensive genetic analysis by next-generation sequencing of 480 patients

Molecular diagnosis of patients with von Willebrand disease is pending in most populations due to the complexity and high cost of conventional molecular analyses. The need for molecular and clinical characterization of von Willebrand disease in Spain prompted the creation of a multicenter project (PCM-EVW-ES) that resulted in the largest prospective cohort study of patients with all types of von Willebrand disease. Molecular analysis of relevant regions of the VWF, including intronic and promoter regions, was achieved in the 556 individuals recruited via the development of a simple, innovative, relatively low-cost protocol based on microfluidic technology and next-generation sequencing. A total of 704 variants (237 different) were identified along VWF, 155 of which had not been previously recorded in the international mutation database. The potential pathogenic effect of these variants was assessed by in silico analysis. Furthermore, four short tandem repeats were analyzed in order to evaluate the ancestral origin of recurrent mutations. The outcome of genetic analysis allowed for the reclassification of 110 patients, identification of 37 asymptomatic carriers (important for genetic counseling) and re-inclusion of 43 patients previously excluded by phenotyping results. In total, 480 patients were definitively diagnosed. Candidate mutations were identified in all patients except 13 type 1 von Willebrand disease, yielding a high genotype-phenotype correlation. Our data reinforce the capital importance and usefulness of genetics in von Willebrand disease diagnostics. The progressive implementation of molecular study as the first-line test for routine diagnosis of this condition will lead to increasingly more personalized and effective care for this patient population

The relationship between characteristics of nurses and their attitude toward nursing diagnosis: a cluster analysis

Aims and objectives To classify a sample of nurses into groups with similar attitude towards nursing diagnosis (ND), using the cluster analysis method, to explore differences in the characteristics between the identified profile groups. The characteristics tested included sociodemographic and professional aspects, and the degree of contact with ND. Methods A cross-sectional explorative design was used in a convenience sample of 548 registered nurses (RNs) was recruited through snowball sampling between the contacts of collaborators of the research group. It was used the Nursing Diagnosis Scale (PND): a 20-item scale that uses the semantic differential method to assess attitudes towards ND. Descriptive statistics were used to summarise data. A hierarchical cluster analysis was employed to categorise the participants into mutually exclusive clusters with similar attitude profiles based on their responses to the 20 items of the PND. Results A three-cluster solution was considered the most suitable. Clusters 1, 2 and 3 comprise RNs with positive, neutral and negative attitudes towards ND, respectively. Conclusion RNs who work in the management field show a better attitude. The increasing awareness of the Spanish managers regarding the benefits of the use of ND in practice could be related to this finding. Contact with ND appears to be a common factor among nurses with positive attitudes; the greater the interaction with ND by nurses, the better the attitudes

A forma abreviada da escala posições frente ao diagnóstico de enfermagem: desenvolvimento e avaliação psicométrica

The Positions on Nursing Diagnosis (PND) is a scale that uses the semantic differential technique to measure nurses' attitudes towards the nursing diagnosis concept. The aim of this study was to develop a shortened form of the Spanish version of this scale and evaluate its psychometric properties and efficiency. A double theoretical-empirical approach was used to obtain a short form of the PND, the PND-7-SV, which would be equivalent to the original. Using a cross-sectional survey design, the reliability (internal consistency and test-retest reliability), construct (exploratory factor analysis, known-groups technique and discriminant validity) and criterion-related validity (concurrent validity), sensitivity to change and efficiency of the PND-7-SV were assessed in a sample of 476 Spanish nursing students. The results endorsed the utility of the PND-7-SV to measure attitudes toward nursing diagnosis in an equivalent manner to the complete form of the scale and in a shorter time.El Position on Nursing Diagnosis (PND) es una escala que utiliza la técnica del diferencial semántico para medir las actitudes hacia el concepto diagnóstico enfermero. El estudio objetivó desarrollar una forma abreviada de la versión española de esta escala, evaluar sus propiedades psicométricas y eficiencia. Se utilizó un doble enfoque empírico-teórico para obtener una forma reducida del PND, el PND-7-SV, que fuera equivalente a la original. Mediante un diseño transversal a través de encuesta, se evaluó la fiabilidad (consistencia interna y fiabilidad test-retest), validez de constructo (análisis factorial exploratorio, técnica de grupos conocidos y validez discriminante) y de criterio (validez concurrente), sensibilidad al cambio y eficiencia del PND-7-SV en una muestra de 476 estudiantes de enfermería españoles. Los resultados avalaron la utilidad del PND-7-SV para medir las actitudes hacia el diagnóstico enfermero de manera equivalente a la forma completa de la escala y en un tiempo más reducido